Unlocking the potential of flavonoids: Natural solutions in the fight against colon cancer.

Colorectal cancer (CRC) is a major cause of cancer-related deaths worldwide, underscoring the importance of understanding the diverse molecular and genetic underpinnings of CRC to improve its diagnosis, prognosis, and treatment. This review delves into the adenoma-carcinoma-metastasis model, emphasizing the "APC-KRAS-TP53" signature events in CRC development. CRC is categorized into four consensus molecular subtypes, each characterized by unique genetic alterations and responses to therapy, illustrating its complexity and heterogeneity. Furthermore, we explore the role of chronic inflammation and the gut microbiome in CRC progression, emphasizing the potential of targeting these factors for prevention and treatment. This review discusses the impact of dietary carcinogens and lifestyle factors and the critical role of early detection in improving outcomes, and also examines conventional chemotherapy options for CRC and associated challenges. There is significant focus on the therapeutic potential of flavonoids for CRC management, discussing various types of flavonoids, their sources, and mechanisms of action, including their antioxidant properties, modulation of cell signaling pathways, and effects on cell cycle and apoptosis. This article presents evidence of the synergistic effects of flavonoids with conventional cancer therapies and their role in modulating the gut microbiome and immune response, thereby offering new avenues for CRC treatment. We conclude by emphasizing the importance of a multidisciplinary approach to CRC research and treatment, incorporating insights from genetic, molecular, and lifestyle factors. Further research is needed on the preventive and therapeutic potential of natural compounds, such as flavonoids, in CRC, underscoring the need for personalized and targeted treatment strategies.

The ethyl acetate extract of Vernonia amygdalina leaf ameliorates gemcitabine effect against migration and invasion of PANC-1 cells via down-regulation the VEGF, COX2, and RAS/MEK pathways.

Individuals diagnosed with cancer often turn to the use of herbal remedies with the intention of treating and ameliorating the condition, impeding the progression of metastasis, enhancing immune function, mitigating stress, and inducing relaxation. Recently, medicinal plants were combined with conventional chemotherapy to decrease the side effects and increase the effectiveness of chemotherapy. This study showed the effectiveness of gemcitabine (Gem) was significantly increased after being used together with ethyl acetate extract obtained from Vernonia amygdalina (Eav) leaves. The combination doses of Eav and Gem were determined based on cytotoxic activity using the MTT assay method. The anticancer effect of this combination was identified by several parameters including the apoptosis effect, anti-migration, and anti-invasion activities of PANC-1 cells. Furthermore, this effect was explained via protein expression evaluation using immunohistochemical and flow cytometry. The Eav has a better Inhibitory Concentration 50 (IC50) than Gem of 21.19 ± 0.64 µg/mL and 164.78 ± 1.40 µg/mL. The combination of Eav and Gem at IC50 (1:1) has the strongest activity than Eav and Gem alone at 500.00 µg/mL. The anti-cancer effect of this combination showed significantly increased levels of apoptosis, particularly in the early phase of 17.46 ± 0.35 % (p < 0.0001) than Eav and Gem alone of 7.76 ± 0.25 % and 7.06 ± 0.20 %. A similar impact was evaluated in the migration and invasion of PANC-1 cells after the combination treatment. The % relative migration and cell invasion were significantly decreased compared to the control group and Eav or Gem alone by 21.49 ± 0.96 % and 125.25 ± 5.25 cells, respectively (p < 0.0001). This study found that signature molecules of VEGF, COX2, RAS, and MEK were down-regulated after treatment. Our study suggested that the Eav ameliorates the Gem effect against PANC-1 cells through apoptosis, migration, and invasion influence via RAS/MEK pathways.

Phyllanthus emblica: a comprehensive review of its phytochemical composition and pharmacological properties.

Phyllanthus emblica Linn, a prominent member of the euphorbiaceae family, exhibits extensive distribution across a multitude of tropical and subtropical nations. Referred to as "Balakka" in Indonesia, this plant assumes various names across regions, such as "kimalaka," "balakka," "metengo," "malaka," and "kemloko" in North Sumatra, Ternate, Sundanese, and Java respectively. Phyllanthus emblica thrives in tropical locales like Indonesia, Malaysia, and Thailand, while also making its presence felt in subtropical regions like India, China, Uzbekistan, and Sri Lanka. The fruits of Balakka are enriched with bioactive constituents recognized for their wide-ranging benefits, including antioxidant, anti-aging, anti-cholesterol, anti-diabetic, immunomodulatory, antipyretic, analgesic, anti-inflammatory, chemoprotective, hepatoprotective, cardioprotective, antimutagenic, and antimicrobial properties. Comprising a spectrum of phenolic compounds (such as tannins, phenolic acids, and flavonoids), alkaloids, phytosterols, terpenoids, organic acids, amino acids, and vitamins, the bioactive components of Malacca fruit offer a diverse array of health-promoting attributes. In light of these insights, this review aims to comprehensively examine the pharmacological activities associated with P. emblica and delve into the intricate composition of its phytochemical constituents.

Effect of Phaleria Macrocarpa (Scheff.) Boerl Leaf Ethanol Extract on Serum IL-6 and TNF-α Levels in Diabetic Rats.

Background: Hyperglycemia conditions in diabetes mellitus (DM) can turn on pro-inflammatory cytokines like IL-6 and TNF-α. These cytokines play a role in insulin resistance and the development of DM complications. People in Indonesia have used Phaleria macrocarpa to treat diabetes, but the leaf of this plant has not been studied to see if it can reduce inflammation. Objective: This study aims to analyze the effect of ethanolic extract of Phaleria macrocarpa leaves (EEPML) in serum IL-6 and TNF-α levels of diabetic rats. Methods: This study was an experiment with a post-test-only control group design. Thirty 8-week-old male Wistar rats were used in the study. They were split into six groups: K1 was the normal control group; K2 was the DM control group; K3, K4, and K5 were given EEPML at doses of 125, 250, and 500 mg/KgBW; and K6 was given metformin 45 mg/KgBW orally once a day for 14 days. A high-fat diet and a 30 mg/KgBWi.p injection of streptozotocin were used to make the diabetic rat model. ELISA method for measuring serum IL-6 and TNF-α levels. The Kruskal-Wallis and the Mann-Whitney test were used to examine the differences between the groups. Results: There were significant differences between treatment groups in the mean levels of serum IL-6 (p=0.017), but there were no significant differences in the mean levels of serum TNF-α (p>0.05). Conclusion: Administration of Phaleria macrocarpa leaf ethanol extract 125 mg/KgBW reduced serum IL-6 levels but could not significantly reduce serum TNF-α levels in diabetic rats.

Anti-Aging Activity of Andaliman (Zanthoxylum Acanthopodium DC) Fruit Ethanol Extract on Brain Weight and p16INK4a Expression of Hippocampus in Aging Model Rats.

Background: Physiological aging and due to oxidative stress in long term will have an impact on cellular response disorders, can caused aging of hippocampus and senility. Brain weight is known to decrease with age and p16INK4a as aging biomarkers have been investigated. Andaliman is one of typical herbal plants from North Sumatra has been widely used as an antioxidant, anti-inflammatory and anti-aging. Objective: This study was evaluated effect of andaliman (Zanthoxylum acanthopodium DC) fruit ethanol extract (AEE) on brain weight and p16INK4a expression in aging model rats. Methods: This study was carried out experimentally of 24 male wistar rats. The treatment group consisted of 4 groups; KN= negatif control (normal), KP= positif control (aging model rat), P1 and P2= aging model rat + AEE at dose 150 and 300mg/kgbw, respectively. The aging model rats were D-galactose-induced at dose of 150mg/kgbw for 8 weeks. Brain weigth were recorded by digital scales. p16INK4a expression using immunohistochemical methods. The data analysis with Anova test. Results: This study showed differences brain weight between groups (p=0.523). Brain weight in P1 (1.34±0,06) and P2 (1.30±0.09) tendency increased than KP (1.29±0.62). The p16INK4a expression between groups significant difference (p=0.041), continued with post hoc Least Significant Difference (LSD) showed p16INK4a expression in KN significant decreased than KP (p=0.027). Likewise, p16INK4a expression in P2 was significant decreased than KP (p=0.010). Conclusion: Andaliman ethanol extract at a dose 300mg/kgbw for 8 weeks was improved aging process caused D-galactose induced.

The anticancer activities of Vernonia amygdalina Delile. Leaves on 4T1 breast cancer cells through phosphoinositide 3-kinase (PI3K) pathway.

Vernonia amygdalina Delile (Asteraceae) is used in traditional medicine to treat diabetes mellitus, and some research provides its activity to treat breast cancer. The aim of this study is to assess the anticancer activity of Vernonia amygdalina Delile leaves fractions on 4T1 breast cancer cells. Analysis of phytochemical compounds were carried out with LC-MS/MS. Cytotoxic activity was determined using the MTT method in the 4T1 cell line. Apoptosis, the cell cycle, and PI3K and mTOR profiles were analyzed with flow cytometry. The phytochemicals found were diterpene (ingenol-3-angelate) and some phenolics (chlorogenic acid and 4-methoxycinnamic acid), flavonoids (apigetrin, apigenin, luteolin, diosmetin, baicalin, rhoifolin, and scutellarin), and coumarines (7-hydroxycoumarine, 4-methylumbelliferone, and 4-methylumbelliferyl glucuronide). The results of the MTT assay showed that the IC50 values n-hexane fraction, ethylacetate fraction (EAF), and ethanol fractions were 1,860.54 ± 93.11, 25.04 ± 0.36, and 1,940.84 ± 96.37 µg/mL, respectively. EAF induced early and late apoptosis, inhibited cell cycle progression on the G2/M phase, and inhibited PI3K and mTOR expression. The EAF of Vernonia amygdalina Delile leaves showed anticancer activity on 4T1 breast cancer cells through induction of apoptosis, enhanced cell accumulation on G2/M phases in the cell cycle, and inhibited expression of PI3K and mTOR.

The PI3KCA and AKT Inhibitory Activities of Litsea Cubeba Lour. Fruits and Heartwoods Towards Hela Cells.

AIM: To investigated the activities of chloroform fractions at pH 7 of Litsea cubeba Lour. Fruits and heartwoods (CF-7F and CF-7H) in decrease expression of PI3KCA, Akt-1 and Akt-2 genes towards cervical cancer cell culture (HeLa) experiments in vitro. MATERIAL AND METHODS: CF-7F and CF-7H (12.5 and 25 µg/mL) were tested for its potential inhibition on gene expression of PI3KCA, Akt-1 and Akt-2 genes by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) method. RESULT: CF-7F and CF-7H were showed the activity to reduce the expression of PI3KCA, Akt-1 and Akt-2 genes. CONCLUSION: Our results suggest that CF-7F and CF-7H significantly inhibit the expression of PI3KCA, Akt-1 and Akt-2 genes.

Cell Cycle Inhibition of Ethylacetate Fraction of Zanthoxylum Acanthopodium DC. Fruit against T47D Cells.

BACKGROUND: The use of medicinal plants is increasing in several decades for relief many diseases. Indonesia consists of thousands of islands with various plants and the manners of the community using plants as a treatment for every disease traditionally. AIM: Cytotoxic activity of ethyl acetate fraction (EAF) of Zanthoxylum acanthopodium fruit was tested towards T47D breast cancer cells. METHODS: The in vitro cytotoxic activity was performed by MTT assay, and the result was expressed as the IC50 (Inhibitory Concentration), and cell cycle inhibition was investigated by flow cytometry to assess the inhibition in every phase of cell cycle, and the role of expression cyclin D1 and p53 in cell cycle inhibition were performed by immunocytochemistry. RESULTS: EAF was showed to have high activity with a value of IC50 48.94 ± 0.32 µg/mL. EAF of 25 µg/mL caused cell accumulation at G0/G1 (60.48%) and in a control cell (51.69%) and decreased expression of cyclin D1 and increased expression of p53. CONCLUSION: The results obtained in this study provided scientific support for further investigation on compounds in Z. acanthopodium fruit which in the future could be used for medication.

The Effect of Puguntano Leaf Extract (Curanga Fel - Terrae Merr.) On P38 Mapk Levels and Glut-4 Expression in Type 2 Diabetic Rat Muscle.

BACKGROUND: Puguntano (Curanga feel-terrae Merr.) contains flavonoids, saponins, tannins, and steroids/ terpenoids which improved post-receptor insulin signalling in rats model of type 2 diabetes mellitus (T2DM). AIM: This study aimed to determine the effect of puguntano leaf extract on p38 mitogen-activated protein kinase (MAPK) levels and glucose transporter-4 (GLUT-4) expression in diabetic rats muscle. METHODS: Forty-eight male Wistar rats had T2DM induced using a combination of feeding a high-fat diet for 5 weeks and multiple intraperitoneal injections of low-dose streptozotocin (30 mg/kg). The diabetic rats were randomly divided into control and treatment groups, and 200 mg/kg/day puguntano extract was administered orally for 10 days to treatment group. Subsequently, p38 MAPK levels were measured by Sandwich Elisa and plasma membrane GLUT-4 expression was evaluated by Immunohistochemistry in their gastrocnemius muscles. RESULTS: There were significantly higher p38 MAPK levels and GLUT-4 expression in the treatment group than in the control group. CONCLUSION: These data suggest that a puguntano leaf extract can improve post-receptor insulin signalling by enhancing p38 MAPK levels and GLUT-4 expression in a rat model of T2DM.

Anticancer Activity of Polyisoprenoids from Avicennia alba Blume. in WiDr Cells.

Colorectal cancer is the third most common cancer world wide and has been occurred more in developing regions. The use of conventional chemotherapy agents may lead to various adverse effects. Therefore, it is required to find the potential drug for anticancer from alternative source of natural product including mangrove plants. The present study was conducted to determine the anticancer activity of polyisoprenoids from Avicennia alba Blume. leaves (PAL) in WiDr cells. Cell cycle inhibition, apoptosis activity, and suppression of cyclooxygenase-2 (COX-2) were also evaluated. The anticancer activity of PAL was determined by observing the activity of these compounds against WiDr cells using the [3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazolium bromida] MTT assay. Inhibition of the cell cycle and increased apoptosis were analysed by flowcytometry. Suppression of COX-2 was analysed using immunocytochemistry. PAL exhibited anticancer activity against WiDr cells with an IC50 of 173.775 µg/mL. Cell cycle analysis revealed that the inhibition occurred in the G0-G1 phase, and apoptosis occurred in the early apoptosis phase. Furthermore, the result of an analysis of COX-2 expression showed that PAL enabled the suppression of COX-2 expression. PAL can be used as anticancer agents against WiDr colon cancer cells. However, in-vivo studies is required to confirm the in-vitro finding of the anticancer activity of polyisoprenoid extract.

Evaluation of Cytotoxic Activity Alkaloid Fractions of Zanthoxylum acanthopodium DC. Fruits.

AIM: This study was carried out to investigate cytotoxic activity towards T47D, 4T1, MCF-7, HeLa, and Raji cells of alkaloid fractions of Zanthoxylum acanthopodium DC. fruits. Zanthoxylum acanthopodium DC. METHODS: The fruit was extracted by maceration. The ethanol extract was fractionated with liquid-liquid extraction using n-hexane, chloroform at pH 3, 7, and 9 to obtained alkaloid fractions. Cytotoxic activity for fraction chloroform at pH 7 and 9 was determined with MTT assay. RESULTS: The IC50 of fraction chloroform at pH 7 and 9 was (92.67 ± 1.37; 71.87 ± 1.04; 159.87 ± 0.63; 123.39 ± 0.81; and 103.09 ± 0.58 µg/mLfor pH 7) and (451.29 ± 25.48; 247.18 ± 2.82; 318.46 ± 5.40; 303.96 ± 8.75; and 181.45 ± 1.35 µg/mL for pH 9) respectively. CONCLUSION: The results reveal that alkaloid fractions at pH 7 and 9 of Zanthoxylum acanthopodium DC. Fruits have cytotoxic activity. Our further study is to isolate and assesses anticancer activity from alkaloid compounds.

Prominent Secondary Metabolites from Selected Genus of Avicennia Leaves.

BACKGROUND: Mangrove plants distributed in the intertidal of the tropical and subtropical region including in North Sumatra, Indonesia. The production of secondary metabolite compounds is well known to mangroves. Characterisation of prominent compounds from mangrove plants such as genus of Avicennia is required to explore for their biological and pharmacological properties of these compounds. AIM: The purpose of this research was to analyse the prominent secondary metabolites through the characterisation of phytochemical, physicochemical, and microscopic of the mangrove genus Avicennia leaves, particularly Avicennia alba, A. lanata, A. marina, and A. officinalis. METHODS: Phytochemical screening was carried out on Avicennia spp leaves to the established process. Physicochemical characters of mangrove leaves were investigated by simplicial powder consisting of moisture content, water-soluble, ethanol-soluble, ash content and ash soluble acid according to the WHO formula. Microscopic analysis on the simplicial powder was carried out based on the WHO procedure. RESULTS: The result showed that physicochemical feature displays diversity among the species and important findings on the water concentration was less than 10% as a prerequisite for the drug. The phytochemical search of simplified grain also depicted divergence among the species, only alkaloid, saponin, and triterpenoid or phytosterol were found entirely in Avicennia spp leaves. Microscopic search found a similar type of stoma in Avicennia spp leaves, namely diacytic. CONCLUSION: The prominent secondary metabolites in Avicennia spp leaves consisting of alkaloid and saponin in simplicial and triterpenoid/sterol was either in simplicial or hexane extract. The present study may provide significant pharmacological properties from mangrove Avicennia genus green foliages, which could accelerate another prospect for non-wood mangrove utilisation.

Cyclin D1, Caspase 9 and P53 Expressions in T47D Cell Lines after Treatment of Plectranthus amboinicus, (Lour.) Spreng. Leaves Ethanolic Extract Nanoparticles.

BACKGROUND: The application of nanotechnology is aimed to enhance the capability of the chemical compounds of Plectranthus amboinicus (Lour.) Spreng. leaves activity. The contact area of the particle surface becomes larger in nano size, which can increase the amount of the isolated active substance. The nanoparticles of this extract exhibited the most effective impact in decreasing the cell number of T47D by induction of apoptosis and arresting the cell cycle. AIM: To evaluate the expressions of cyclin D1, caspase-9 and p53 in T47D cell lines treated by Plectranthus amboinicus, (Lour.) Spreng. Leaves Ethanolic Extract Nanoparticles (PAEEN). METHODS: The inhibition of cyclin D1 was done by flow cytometry assay. The expression of cyclin D1, caspase-9 and p53 were observed by immunocytochemistry assay. RESULTS: Plectranthus amboinicus, (Lour.) Spreng. Leaves Ethanolic Extract Nanoparticles (PAEEN) on concentration IC50 (89.2 µg/mL) inhibited the expression of cyclin D1 around 5.59%. Further, the immunocytochemistry assay indicated that there was an inhibition of cyclin D1, upregulation of caspase-9 and restoration of p53, indicated by their expression. CONCLUSIONS: The Plectranthus amboinicus, (Lour.) Spreng. Leaves Ethanolic Extract Nanoparticles could be an effective anticancer by improving its downregulation on cyclinD1, upregulation caspase-9 and p53.

Effect of Salt and Fresh Water Concentration on Polyisoprenoid Content in Bruguiera cylindrica Seedlings.

BACKGROUND: Mangrove forest is a typical forest found along the coast or river mouth which is affected by tides and salinity. Although polyisoprenoid was widespread in the plant kingdom, the physiological roles of these compounds are not well understood, especially from mangrove plants. It is therefore essential to characterize the polyisoprenoid content under abiotic stress. AIM: This study aimed to determine the effect of salinity and subsequent fresh water change on polyisoprenoids concentration in Bruguiera cylindrica seedlings. METHODS: Bruguiera cylindrica planted in a greenhouse for three months under various salinity concentrations. After three months grew under variable salinity, these seedlings were then divided into two treatment groups, and grown for another three months: one continuously in a salt solution and another in fresh water to relieve salt stress. The leaves and roots of B. cylindrica seedlings were harvested after six months of cultivation. The leaves and roots of B. cylindrica seedlings were extracted for polyisoprenoids content and composition analyzed using two-dimensional thin layer chromatography. RESULTS: Polyisoprenoids composition under salinity and subsequent fresh water with dominating dolichols (more than 90%) were found in leaves and roots of B. cylindrica seedlings referring type I of polyisoprenoid composition. The carbon chain length of dolichols located in the leaves and roots were ranging from C75-C100 and C75-C105, respectively. CONCLUSON: Dolichol dominated over polyprenol both in B. cylindrical leaves and roots under salinity and subsequent relief supported the previous finding on the predominance dolichols over polyprenols in mangrove plants. The present study suggested the significance of dolichols in the adaptation to cope with salt stress and or water stress.

Cell Cycle Inhibition and Apoptotic Induction of Vernonia amygdalina Del. Leaves Extract on MCF-7 Cell Line.

AIM: The research aimed to determine the cytotoxic activity, cell cycle inhibition, and apoptosis induction of the ethyl acetate extract of the African leaves Vernonia amygdalina Del. on the MCF-7 cancer cells. METHODS: The extraction of Vernonia amygdalina Del. leaves was done using the maceration method whereas the cytotoxic was performed using MTT assay. After that, the cell cycle testing and apoptosis induction were conducted using flow cytometry assay. RESULTS: The IC50 values of n-hexane, ethyl acetate, and ethanol extract of Vernonia amygdalina Del. on the MCF-7 cancer cells were 206.211 ± 0.99, 50,365 ± 0.07, and 967.033 ± 2.68 µg/mL, respectively. The percentage of the cycle cell results in the G0-G1 phase in the cell control with 72.08% decreased in the treatment with ethyl acetate extract 1/2 IC50 with 62.58% and 1/5 IC50 with 44.72%. For the S and G2-M phase, the highest percentage was found in the ethyl acetate extract 1/5 IC50 treatment with 47.27% and 9.50% which were higher than the control cells with 23.26% and 5.90%. CONCLUSION: Based on the results, the Vernonia amygdalina Del. extract provides chemopreventive agent as anti-cancer. Our future study will assess the mechanism of ethyl acetate fraction in inhibiting angiogenesis and metastatic in breast cancer.

Cytoprotective Activity of Ethylacetate Fraction of Picria fel-terrae Lour. Herbs.

AIM: Free radicals produce from metabolism or enviromental which interact continously with biological system. Picria fel-terrae Lour. herbs have been used as antioxidant and treat various diseases. The aim of this study was to evaluate cytoprotective activity of ethylacetate fraction (EAF) of Picria fel-terrae Lour. herbs. METHODS: Cytoprotective activity were determined by MTT asay and flow cytometry assay on Vero cells which induced with H2O2 0.8 mM. RESULTS: EAF at 100 µg/mL were showed highest viability (88.83 ± 2.90%) and ROS expression (66.75%) on Vero cells. CONCLUSION: EAF of Picria fel-terrae Lour. herbs have cytoprotective activity.

Antimigration Activity of an Ethylacetate Fraction of Zanthoxylum acanthopodium DC. Fruits in 4T1 Breast Cancer Cells

Objective: This study was carried out to investigate the antimigration activity of Zanthoxylum acanthopodium DC. in the 4T1 breast cancer cell line. Methods: Zanthoxylum acanthopodium DC. fruit powder was extracted by maceration method with n-hexane and ethylacetate solvents. Cytotoxicity and proliferation were assessed using the MTT method and the cell cycle by flow cytometry. In addition, wound healing assays were conducted by a microscopic method, and expression of COX-2 and VEGFR-2 were determined using qRT-PCR. Results: The IC50 of the ethylacetate fraction (EAF) was 48.1 ± 1.06 µg/mL. The EAE at a concentration 10 µg/mL with viable cells was 62.3 ± 0.28% after 72 h incubation, with accumulation in the G2-M phase, inhibition of cell migration in the wound healing assay, and decrease in expression of COX-2 (0.62 ± 0.01) and VEGFR-2 (0.39 ± 0.003). Conclusion: The results reveal that an ethylacetate fraction of Zanthoxylum acanthopodium DC. fruits provides effective antimigration effects. Further studies are now planned to assess the potential of the ethylacetate fraction to inhibit angiogenesis in breast cancer and determine underlying mechanisms.